Combining protein design with cell engineering to unlock the future of curative cell therapies

Part of the Protein and Cell Engineering group at Lyell, which applies cutting-edge protein design, high-throughput molecular biology, and next-gen sequencing to engineer CAR T cells with new functions that enable them to be effective against solid tumors. • Culture of suspension and adherent cells, both human cell lines and human primary T Cells• Viral vector transduction, characterization and analysis through multi-color flow cytometry, cell sorting, MSD, and FlowJo analysis • Co-culture killing assays via the Incucyte platform• High throughput viral vector production

Development of transgenic murine embryonic stem cell lines and human cancer cell lines using an array of molecular biology methods, from generating novel molecular tools for cell engineering, to the generation of transgenic embryonic stem cell lines, and characterization and genotyping of newly established cell lines.• Developing site-specific nucleases (CRISPR/Cas9) for genetic engineering of murine embryonic stem cells• Transfection and culture of murine ESCs and human immortalized cell lines• Characterizing and identifying clones with correctly targeted transgenes through PCR based genotyping and functional characterization with RT-qPCR• Vector subcloning to create DNA for homologous recombination, using a variety of current techniques including Gibson assembly and other restriction enzyme free cloning strategies• Culture of mammalian cells for imaging and other molecular biology assays• Utilizing the auxin-degron system to examine short term disruptions to the nucleolus • Western blotting using the LI-COR imaging system

Utilizing an array of molecular biology techniques in the development of transgenic human embryonic stem cell lines, from generating novel molecular tools in the cell engineering process to characterization and genotyping of established cell lines.• Developing site-specific nucleases (CRISPR/Cas9 and TALENs) for genetic engineering of hESCs• Producing lentivirus and retrovirus for knockdown, overexpression, and lineage tracing• Identifying clones with correctly targeted transgenes• Extensive work in vector subcloning to create homologous recombination donors, using a variety of current techniques including inFusion and Golden Gate cloning• Culture of mammalian cells for testing and evaluating molecular tools

Development of viral vectors, from plasmid engineering to the generation of recombinant viruses containing optogenetic payloads, and application of those vectors in NHPs. • Single unit electrophysiology and optogenetics • Mammalian cell culture and recombinant virus production (rAAV and lentivirus)• Behavioral training and survival surgeries with NHPs• Writing IRB and IACUC applications• Lab management, including developing SOPs, maintaining laboratory equipment, purchasing and inventory, and Environmental Health and Safety compliance

Investigating the paraoxonase family of proteins and their relationship to Central Nervous System cell lines while writing and defending a thesis from this research• Tissue culture of murine primary cells and cell lines • Western blotting • Enzymatic activity assays

Assisting medical technicians in a biochemical genetics laboratory• Preparing chemical reagents• Maintaining patient records• Laboratory maintenance

Assisting a small animal veterinarian in a private practice• Animal handling, surgery and anesthesia, and radiology• Administering injections/medication, calculating dosages, and filling prescriptions• Discussing care options with clients• General office management, scheduling, and maintaining records• Care for in-house animals, including administering medications and feeding